Intracellular mannitol, a product of glucose metabolism in staphylococci

Edwards, K.G.; Blumenthal, Harold J.; Khan, Mohammad Naseem; Slodki, Morey E.

doi:10.1128/jb.146.3.1020-1029.1981

Cited by 18 publications

(12 citation statements)

References 32 publications

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“…We verified that the production of Man‐ol‐1‐ P and Man‐ol was not an artefact resulting from the use of nongrowing cells and high cell densities during the in vivo NMR experiments, as intracellular Man‐ol‐1‐ P was also detected in growing cells of the LDH d strain (≈ 15 m m during the late exponential growth phase). To our knowledge, this is the first report of the production of Man‐ol and Man‐ol‐1‐ P by L. lactis , although these products have been earlier observed in other bacteria, such as Staphylococcus aureus , Streptococcus mutans , and Escherichia coli [28–32]. Recent work with a different construct of LDH d showed an alternative strategy for NAD + regeneration in which acetate is reduced to ethanol [3].…”

Section: Discussionmentioning

confidence: 99%

Metabolic characterization of Lactococcus lactis deficient in lactate dehydrogenase using in vivo¹³C‐NMR

Neves

Ramos

Shearman

et al. 2000

European Journal of Biochemistry

104

112

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The metabolism of glucose by nongrowing cells of Lactococcus lactis strain FI7851, constructed from the wild-type L. lactis strain MG1363 by disruption of the lactate dehydrogenase (ldh) gene [Gasson, M.J., Benson, K., Swindel, S. & Griffin, H. (1996) Lait 76, 33±40] was studied in a noninvasive manner by 13 C-NMR. The kinetics of the build-up and consumption of the pools of intracellular intermediates mannitol 1-phosphate, fructose 1,6-bisphosphate, 3-phosphoglycerate, and phosphoenolpyruvate as well as the utilization of [1-13 C]glucose and formation of products (lactate, acetate, mannitol, ethanol, acetoin, 2,3-butanediol) were monitored in vivo with a time resolution of 30 s. The metabolism of glucose by the parental wild-type strain was also examined for comparison. A clear shift from typical homolactic fermentation (parental strain) to a mixed acid fermentation (lactate dehdydrogenase deficient; LDH d strain) was observed. Furthermore, high levels of mannitol were transiently produced and metabolized once glucose was depleted. Mannitol 1-phosphate accumulated intracellularly up to 76 mm concentration. Mannitol was formed from fructose 6-phosphate by the combined action of mannitol-1-phosphate dehydrogenase and phosphatase. The results show that the formation of mannitol 1-phosphate by the LDH d strain during glucose catabolism is a consequence of impairment in NADH oxidation caused by a highly reduced LDH activity, the transient production of mannitol 1-phosphate serving as a regeneration pathway for NAD 1 regeneration. Oxygen availability caused a drastic change in the pattern of intermediates and end-products, reinforcing the key-role of the fulfilment of the redox balance. The flux control coefficients for the step catalysed by mannitol-1-phosphate dehydrogenase were calculated and the implications in the design of metabolic engineering strategies are discussed.Keywords: redox balance; noninvasive 13 C-NMR; Lactococcus lactis; biosynthesis of mannitol; mannitol-1-phosphate dehydrogenase.Lactic acid bacteria play a central role in the dairy industry because of their widespread use as starter cultures in milk fermentation. The relatively simple metabolic strategy of these organisms that utilize sugars primarily to generate energy and not for growth, makes them an attractive model system for studies aiming at the construction of improved strains by metabolic engineering. However, a deep understanding of the metabolic network as well as of the interdependence relationships among the different steps are essential to establish a rational and systematic methodology that will enable successful manipulation of carbohydrate metabolism.Recent work aiming at the metabolic characterization of lactic acid bacteria revealed several strategies for regeneration of NAD 1 during the metabolism of carbohydrates. For instance, Oenococcus oeni produces erythritol to consume the reduced coenzymes formed in the glycolytic pathway [1]; a strain of Lactobacillus plantarum deficient in both l-and d-lactate dehydrogenases produce...

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Section: Discussionmentioning

confidence: 99%

Metabolic characterization of Lactococcus lactis deficient in lactate dehydrogenase using in vivo¹³C‐NMR

Neves

Ramos

Shearman

et al. 2000

European Journal of Biochemistry

104

112

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“…The accumulation of mannitol as an intracellular product of glucose metabolism has been reported in staphylococci, a gram‐positive coccus [20,21]. In heterofermentative lactic acid species such as Lactobacillus brevis (an organism devoid of phosphate acetyl transferase and alcohol dehydrogenase), a mannitol dehydrogenase catalyses the reduction of fructose to yield mannitol [22,23].…”

Section: Discussionmentioning

confidence: 99%

³¹P and ¹³C nuclear magnetic resonance studies of metabolic pathways in Pasteurella multocida

Rager

Binet

Bouvet

1999

European Journal of Biochemistry

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Glucose metabolism of Pasteurella multocida was examined in resting cells in vivo using 13 C NMR spectroscopy, in cell-free extracts in vitro using 31 P NMR spectroscopy and using enzyme assays. The NMR data indicate that glucose is converted by the Embden±Meyerhof and pentose phosphate pathways. The P. multocida fructose 6-phosphate phosphotransferase activity (the key enzyme of the Embden±Meyerhof pathway) was similar to that of Escherichia coli. Nevertheless, and in contrast to that of E. coli, its activity was inhibited by a glycerophosphate. This inhibition is consistent with the very low fructose 6-phosphate phosphotransferase activity found in cell-free extracts of P. multocida using a spectrophotometric method. The dominant end products of glucose metabolism were mannitol, acetate and succinate. Under anaerobic conditions, P. multocida was able to constitutively produce mannitol from glucose, mannose, fructose, sucrose, glucose 6-phosphate and fructose 6-phosphate. We propose a new metabolic pathway in P. multocida where fructose 6-phosphate is reduced to mannitol 1-phosphate by fructose 6-phosphate reductase. Mannitol 1-phosphate produced is then converted to mannitol by mannitol 1-phosphatase.

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“…Endogenously produced mannitol-1-phosphate serving as an inducer of the mtl operon could thus explain the presence of the specific enzymes for its metabolism in uninduced cells. The occurrence of mannitol-1-phosphate or of mannitol in cells grown in the absence of this sugar has also been reported in other microorganisms (3,7). In this report, we deal with the production and turnover of mannitol-1-phosphate in wild strains of E. coli and in some mutants affected in the enzymes involved in mannitol metabolism.…”

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confidence: 68%

Rapid turnover of mannitol-1-phosphate in Escherichia coli

Rosenberg¹,

Pearce²,

Hardy³

et al. 1984

J Bacteriol

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The phosphate moiety of D-mannitol-1-phosphate in Escherichia coli is subject to rapid turnover and is in close equilibrium with Pi and the phosphorus of fructose-1,6-bisphosphate. These three compounds account for the bulk of 32P label found in cells after several minutes of uptake of 32Pi and mannitol-1-phosphate represents some 30% of this label. Mannitol-1-phosphate occurs in E. coli grown on a variety of carbon sources, in the absence of D-mannitol, and is synthesized de novo even in mutants lacking mannitol-1-phosphate dehydrogenase. The mannitol moiety of mannitol-1-phosphate was not affected during the total chase of the P moiety, which exchanged with a half-life of about 30 s. These findings suggest that the rapid equilibration of the phosphorus is a function of an enzyme, possibly a component of the phosphotransferase system, capable of forming a complex that allows the exchange of the phosphate without the equilibration of the mannitol moiety with free mannitol.

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Intracellular mannitol, a product of glucose metabolism in staphylococci

Cited by 18 publications

References 32 publications

Metabolic characterization of Lactococcus lactis deficient in lactate dehydrogenase using in vivo¹³C‐NMR

Metabolic characterization of Lactococcus lactis deficient in lactate dehydrogenase using in vivo¹³C‐NMR

³¹P and ¹³C nuclear magnetic resonance studies of metabolic pathways in Pasteurella multocida

Rapid turnover of mannitol-1-phosphate in Escherichia coli

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Intracellular mannitol, a product of glucose metabolism in staphylococci

Cited by 18 publications

References 32 publications

Metabolic characterization of Lactococcus lactis deficient in lactate dehydrogenase using in vivo13C‐NMR

Metabolic characterization of Lactococcus lactis deficient in lactate dehydrogenase using in vivo13C‐NMR

31P and 13C nuclear magnetic resonance studies of metabolic pathways in Pasteurella multocida

Rapid turnover of mannitol-1-phosphate in Escherichia coli

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Metabolic characterization of Lactococcus lactis deficient in lactate dehydrogenase using in vivo¹³C‐NMR

Metabolic characterization of Lactococcus lactis deficient in lactate dehydrogenase using in vivo¹³C‐NMR

³¹P and ¹³C nuclear magnetic resonance studies of metabolic pathways in Pasteurella multocida